Nuclease‐independent functions of RAG1 direct distinct DNA damage responses in B cells

Abstract

Developing B cells generate DNA double‐stranded breaks (DSBs) to assemble immunoglobulin receptor (Ig) genes necessary for the expression of a mature B cell receptor. These physiologic DSBs are made by the RAG endonuclease, which is comprised of the RAG1 and RAG2 proteins. In pre‐B cells, RAG‐mediated DSBs activate the ATM kinase to coordinate canonical and non‐canonical DNA damage responses (DDR) that trigger DSB repair and B cell developmental signals, respectively. Whether this broad cellular response is distinctive to RAG DSBs is poorly understood. To delineate the factors that direct DDR signaling in B cells, we express a tetracycline‐inducible Cas9 nuclease in Rag1‐deficient pre‐B cells. Both RAG‐ and Cas9‐mediated DSBs at Ig genes activate canonical DDR. In contrast, RAG DSBs, but not Cas9 DSBs, induce the non‐canonical DDR‐dependent developmental program. This unique response to RAG DSBs is, in part, regulated by non‐core regions of RAG1. Thus, B cells trigger distinct cellular responses to RAG DSBs through unique properties of the RAG endonuclease that promotes activation of B cell developmental programs.

Document Details

Document Type
Pub Defense Publication
Publication Date
Nov 16, 2022
Source ID
10.15252/embr.202255429

Entities

People

  • Abby M Green
  • Brendan Mathias
  • Haley A. Schmidt
  • Jeffrey J Bednarski
  • Lynn S White
  • Nima Mosammaparast
  • Rachel Johnston
  • Stephanie J. Crowley

Organizations

  • Alvin J. Siteman Cancer Center
  • American Cancer Society
  • American Society of Hematology
  • Foundation for Barnes-Jewish Hospital
  • Gabrielle’s Angel Foundation for Cancer Research
  • National Cancer Institute
  • National Institute of Allergy and Infectious Diseases
  • St. Louis Children's Hospital
  • United States Department of Defense
  • Washington University in St. Louis

Tags

Fields of Study

  • Biology

Readers

  • Materials Science.
  • Molecular Biology and Genetics